GSM6729140: Sample_SARS-CoV-2_2-24h; Homo sapiens; miRNA-Seq
Total RNA was extracted from Calu-3 infected and non-infected cells at the 24 h, 48 h and 72 h time points, using RNA Easy kit (Qiagen, Cat. #74106, Toronto, Ontario, Canada) following the manufacturer's recommendations. All RNA samples were treated with DNase I, quantified with the NanoDrop™ 2000 Spectrophotometer (Thermo Scientific™, Cat #ND-2000) and kept at -80°C for subsequent experiments Total RNA of each sample was used to prepare the miRNA sequencing library which included the following steps: 1) 3'-adapter ligation with T4 RNA ligase 2 (truncated); 2) 5'-adapter ligation with T4 RNA ligase; 3) cDNA synthesis with RT primer; 4) PCR amplification; 5) extraction and purification of ~130-150 bp PCR amplified fragments (correspond to ~8-15 nt small RNAs) from the PAGE gel. After the completed libraries were quantified with Agilent 2100 Bioanalyzer, the DNA fragments in the libraries were denatured with 0.1M NaOH to generate single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ and finally sequenced for 50 cycles on Illumina Nextseq according to the manufacturer's instruction RNA 16-30nt library